摘要
目的研究在白细胞介素1 (IL-1)及转化生长因子β(TGF-β)影响下,人膝关节软骨细胞中的金属蛋白酶组织抑制因子4 (TIMP-4)蛋白表达水平的变化,从分子水平上研究骨性关节炎(OA)的调控机制,为临床治疗提供理论依据。方法软骨细胞经原代培养、纯化和传代后,除对照组外,分别与IL-1及TGF-β共培养实施细胞因子干预,最后经免疫印迹试验测量TIMP-4蛋白表达水平,各组数据釆用单因素方差分析(ANOVA),以正常软骨对照组均值为参考基数(1.00)。结果在没有细胞因子干预的情况下,软骨细胞均不同程度地表达TIMP-4蛋白, OA患者的软骨细胞表达水平更高,差异有统计学意义(P <0.05);在TGF-β和IL-1干预下, OA患者软骨细胞无论较未经干预的OA软骨细胞,还是经干预的正常软骨细胞, TIMP-4蛋白表达量均呈现统计学差异(P <0.05),而且TGF-β(1.45±0.05)比IL-1 (1.18±0.01)干预效果更明显。结论人原发性膝关节骨性关节炎的软骨细胞中TIMP-4的表达增加,而且可由细胞因子IL-1及TGF-β进行诱导。
Objective To study the changes in the expression level of tissue inhibitor of metalloproteinase-4(TIMP-4) protein in human knee articular chondrocytes under the influence of interleukin-1(IL-1) and transforming growth factor β(TGF-β), and to study the regulatory mechanism of osteoarthritis(OA) at the molecular level, so as to provide a theoretical basis for clinical treatment. Methods After primary culture, purification and passage of chondrocytes, besides the control group, the chondrocytes were respectively co-cultured with IL-1 and TGF-β to implement cytokine intervention. Finally, the expression level of TIMP-4 protein was measured in the Western Blot test.The data of each group were analyzed by one-way analysis of variance(ANOVA), with the mean value of normal cartilage control group as the reference base(1.00). Results In the absence of cytokine intervention, TIMP-4 protein was expressed in chondrocytes to varying degrees, and the expression level of chondrocytes in OA patients was higher, the difference was statistically significant(P <0.05). Under the intervention of TGF-β and IL-1, the expression of TIMP-4 protein in chondrocytes of OA patients showed statistically significant difference(P <0.05), no matter compared with OA chondrocytes without intervention or compared with normal chondrocytes after intervention. The intervention effect of TGF-β(1.45 ± 0.05) was more obvious than that of IL-1(1.18 ± 0.01). Conclusions TIMP-4 in chondrocytes of human primary knee osteoarthritis increases its expression and can be induced by cytokines IL-1 and TGF-β.
引文
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